Decrease in Abundance of apurinic/apyrimidinic Endonuclease
Causes Failure of Base Excision Repair in Culture-Adapted Human
Embryonic Stem Cells

J 2013

Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells

KRUTÁ, Miriama; Lukáš BÁLEK; Renata HEJNOVÁ; Zuzana KUNICKÁ; Lívia EISELLEOVÁ et al.

Základní údaje

Originální název

Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells

Autoři

KRUTÁ, Miriama; Lukáš BÁLEK; Renata HEJNOVÁ; Zuzana KUNICKÁ; Lívia EISELLEOVÁ; Kamil MATULKA; Tomáš BÁRTA; Petr FOJTÍK; Jiří FAJKUS ; Aleš HAMPL; Petr DVOŘÁK a Vladimír ROTREKL

Vydání

Stem Cells, UNITED STATES, WILEY-BLACKWELL, 2013, 1066-5099

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

Genetika a molekulární biologie

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 7.133

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14110/13:00065577

Organizační jednotka

Lékařská fakulta

DOI

https://doi.org/10.1002/stem.1312

UT WoS

000316624300008

Klíčová slova anglicky

APE1; base excision repair; human embryonic stem cells; culture adaptation; genome instability

Štítky

podil

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 25. 8. 2014 15:09, Olga Křížová

Anotace

V originále

The inevitable accumulation of chromosomal abnormalities in human embryonic stem cells (hESCs) during in vitro expansion represents a considerable obstacle for cell replacement therapies. To determine the source of chromosomal abnormalities, we examined hESCs maintained in culture for over 55 months for defects in telomere maintenance and DNA repair. Although prolonged culture affected neither telomerase activity nor nonhomologous end joining, the efficiency of base excision repair (BER) was significantly decreased and correlated with reduced expression of apurinic/apyrimidinic endonuclease 1 (APE1), the major nuclease required for BER. Interestingly, the expression of other BER enzymes was unchanged. Addition of human recombinant APE1 protein to nuclear extracts from late passage hESCs increased BER efficiency to the level typical of early passage hESCs. The link between BER and double-strand breaks (DSB) was demonstrated by decreased DSB release after downregulation of APE1 in early passage hESCs via siRNA. Correspondingly lower APE1 level in late passage hESC resulted in slower and less intensive but long lasting DSB release upon ionizing radiation (IR). Downregulation of APE1 in early passage hESCs also led to approximately 30% decrease in -H2AX signaling following IR, similar to that in late passage hESCs. We suggest that downregulation of APE1 significantly contributes to the failure of BER during long-term culture of hESCs, and further that BER failure is one of the factors affecting the genomic instability of hESCs by altering BER-dependent DSB release and cell cycle/checkpoint signaling.

Návaznosti

GBP302/12/G157, projekt VaV

Název: Dynamika a organizace chromosomů během buněčného cyklu a při diferenciaci v normě a patologii

Investor: Grantová agentura ČR, Dynamika a organizace chromosomů během buněčného cyklu a při diferenciaci v normě a patologii

MSM0021622430, záměr

Název: Funkční a molekulární charakteristiky nádorových a normálních kmenových buněk - identifikace cílů pro nová terapeutika a terapeutické strategie

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Funkční a molekulární charakteristiky nádorových a normálních kmenových buněk - identifikace cílů pro nová terapeutika a terapeutické strategie

NS10237, projekt VaV

Název: Identifikace mechanizmů vzniku chromozomálních aberací při dlouhodobé kultivaci lidských embryonálních buněk in vitro

Investor: Ministerstvo zdravotnictví ČR, Identifikace mechanizmů vzniku chromozomálních aberací při dlouhodobé kultivaci lidských embryonálních buněk in vitro

Přiložené soubory

ZVV_2013_027_1081531_Decrease_in_Abundance.pdf

Požádat o autorskou verzi souboru

Kruta.pdf

Požádat o autorskou verzi souboru

Citovat

KRUTÁ, Miriama; Lukáš BÁLEK; Renata HEJNOVÁ; Zuzana KUNICKÁ; Lívia EISELLEOVÁ; Kamil MATULKA; Tomáš BÁRTA; Petr FOJTÍK; Jiří FAJKUS; Aleš HAMPL; Petr DVOŘÁK a Vladimír ROTREKL. Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells. Stem Cells. UNITED STATES: WILEY-BLACKWELL, 2013, roč. 31, č. 4, s. 693-702. ISSN 1066-5099. Dostupné z: https://doi.org/10.1002/stem.1312.

@article{1081531,
   author = {Krutá, Miriama and Bálek, Lukáš and Hejnová, Renata and Kunická, Zuzana and Eiselleová, Lívia and Matulka, Kamil and Bárta, Tomáš and Fojtík, Petr and Fajkus, Jiří and Hampl, Aleš and Dvořák, Petr and Rotrekl, Vladimír},
   article_location = {UNITED STATES},
   article_number = {4},
   doi = {https://doi.org/10.1002/stem.1312},
   keywords = {APE1; base excision repair; human embryonic stem cells; culture adaptation; genome instability},
   language = {eng},
   issn = {1066-5099},
   journal = {Stem Cells},
   note = {Projekt LSHG-CT-2006-018739 (PLATFORMS FOR BIOMEDICAL DISCOVERY WITH HUMAN ES CELLS) za ČR: AV ČR},
   title = {Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells},
   url = {http://onlinelibrary.wiley.com/doi/10.1002/stem.1312/citedby},
   volume = {31},
   year = {2013}
}

TY  - JOUR
ID  - 1081531
AU  - Krutá, Miriama - Bálek, Lukáš - Hejnová, Renata - Kunická, Zuzana - Eiselleová, Lívia - Matulka, Kamil - Bárta, Tomáš - Fojtík, Petr - Fajkus, Jiří - Hampl, Aleš - Dvořák, Petr - Rotrekl, Vladimír
PY  - 2013
TI  - Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells
JF  - Stem Cells
VL  - 31
IS  - 4
SP  - 693-702
EP  - 693-702
PB  - WILEY-BLACKWELL
SN  - 10665099
N1  - Projekt LSHG-CT-2006-018739 (PLATFORMS FOR BIOMEDICAL DISCOVERY WITH HUMAN ES CELLS) za ČR: AV ČR
KW  - APE1
KW  - base excision repair
KW  - human embryonic stem cells
KW  - culture adaptation
KW  - genome instability
UR  - http://onlinelibrary.wiley.com/doi/10.1002/stem.1312/citedby
N2  - The inevitable accumulation of chromosomal abnormalities in human embryonic stem cells (hESCs) during in vitro expansion represents a considerable obstacle for cell replacement therapies. To determine the source of chromosomal abnormalities, we examined hESCs maintained in culture for over 55 months for defects in telomere maintenance and DNA repair. Although prolonged culture affected neither telomerase activity nor nonhomologous end joining, the efficiency of base excision repair (BER) was significantly decreased and correlated with reduced expression of apurinic/apyrimidinic endonuclease 1 (APE1), the major nuclease required for BER. Interestingly, the expression of other BER enzymes was unchanged. Addition of human recombinant APE1 protein to nuclear extracts from late passage hESCs increased BER efficiency to the level typical of early passage hESCs. The link between BER and double-strand breaks (DSB) was demonstrated by decreased DSB release after downregulation of APE1 in early passage hESCs via siRNA. Correspondingly lower APE1 level in late passage hESC resulted in slower and less intensive but long lasting DSB release upon ionizing radiation (IR). Downregulation of APE1 in early passage hESCs also led to approximately 30% decrease in -H2AX signaling following IR, similar to that in late passage hESCs. We suggest that downregulation of APE1 significantly contributes to the failure of BER during long-term culture of hESCs, and further that BER failure is one of the factors affecting the genomic instability of hESCs by altering BER-dependent DSB release and cell cycle/checkpoint signaling.
ER  -

KRUTÁ, Miriama; Lukáš BÁLEK; Renata HEJNOVÁ; Zuzana KUNICKÁ; Lívia EISELLEOVÁ; Kamil MATULKA; Tomáš BÁRTA; Petr FOJTÍK; Jiří FAJKUS; Aleš HAMPL; Petr DVOŘÁK a Vladimír ROTREKL. Decrease in Abundance of apurinic/apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture-Adapted Human Embryonic Stem Cells. \textit{Stem Cells}. UNITED STATES: WILEY-BLACKWELL, 2013, roč.~31, č.~4, s.~693-702. ISSN~1066-5099. Dostupné z: https://doi.org/10.1002/stem.1312.

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